Species- and age-dependent distribution of retinol and α-tocopherol in the Canidae family during the cold season

Vitamins A and E are known as nutrients involved in metabolic processes and facilitating the adaptation of animals. The metabolism of these vitamins is closely related to lipid metabolism, which has seasonal variations in animals inhabiting northern latitudes. The aim of this study was to investigate the concentrations of retinol (vitamin A) and α-tocopherol (vitamin E) in the cold season (October to February) in tissues of Canidae species that have different ecological characteristics: the blue fox (Vulpes syn. Alopex lagopus), silver fox (Vulpes vulpes), their interspecific hybrids (Vulpes×Vulpes), raccoon dog (Nyctereutes procyonoides), and grey wolf (Canis lupus). In addition, age-related differences in the levels of these vitamins were determined. Results showed the distribution of retinol and α-tocopherol in the body of the canids was similar, with larger amounts being found in the liver and kidney cortex, which play a regulatory role in the metabolism of these nutrients. The higher retinol level in the liver of wolves and interspecific hybrids of blue fox and silver fox, as compared to the retinol level in the liver of other species, might be species-specific and could also be attributed to dietary differences. The blue fox and raccoon dog both have a higher capacity for α-tocopherol accumulation in the cold season, apparently due to their ability to store substantial fat reserves. Retinol and α-tocopherol levels were generally higher in the tissues of mature animals.

Molecular Identification and Antibacterial Activity Analysis of Blue Fox (Vulpes lagopus) β-Defensins 108 and 122

The blue fox (Vulpes lagopus), a fur-bearing animal, is an important component of the breeding industry in China. Semen quality is a key factor for the reproductive process and the breeding effectiveness of the farmed blue fox. However, bacterial contamination in semen samples utilized in artificial fertilization is very common. The β-defensins, a class of important antimicrobial peptides in mammals, could protect the reproductive system of male animals from bacterial invasion, maintain the stability of the genital tract microenvironment and improve semen quality. In this study, molecular cloning and bioinformatics analysis were conducted to analyze the protein structure and function of blue fox β-defensin 108 (Vulpes lagopus beta-defensin 108, vBD108) and 122 (Vulpes lagopus beta-defensin 122, vBD122). To evaluate the bacteriostatic effect of recombinant vBDs (Vulpes lagopus beta-defensins) protein, varying concentrations (0, 25, 50, 100, 200 µg/mL) were taken to evaluate the effects on Escherichia coli and Staphylococcus aureus at different times (0, 2, 4, 6, 8, 12 h). The results showed that vBD108 and vBD122 existed in different forms in protein structure and had antibacterial activity. Both proteins, at 50 µg/mL, had efficacious bacteriostatic activity. This study shows that recombinant vBD108 and vBD122 proteins have good antibacterial activity in vitro. This implies a potential role in improving semen quality and hygienic measures in the process of artificial insemination as an extender of semen dilution with antibacterial activity.

A Retrospective Survey of the Abortion Outbreak Event Caused by Brucellosis at a Blue Fox Breeding Farm in Heilongjiang Province, China

Brucellosis is a common zoonosis in China, resulting in abortion in animals. Outbreaks of abortion in blue foxes caused by Brucella infection have rarely been reported. In the present study, 3–5 mL blood samples collected from the femoral veins of 10 abortuses of blue foxes were assessed by RBPT (Rose Bengal plate test) and SAT (serum tube agglutination test) to preliminarily investigate the source of infection for the clustering of abortion events at a blue fox farm in Heilongjiang Province. Screening experiments showed that all 10 blood samples were positive in the RBPT, while only eight blood samples out of the 10 were positive in the SAT. Subsequently, 10 tissue samples (spleen, lungs, stomach contents, and afterbirth) from the same 10 foxes were assessed using AMOS (acronym for B. abortus, melitensis, ovis, and suis)-PCR (polymerase chain reaction), and sequencing analysis was performed on amplification products to verify the results of the serology survey. Results showed a spectral band of ~731 bp in these samples. BLAST showed sequences of AMOS-PCR products in this study to be 100% similar (E = 0.0) to sequences in B. melitensis strain from GenBank. These data preliminarily indicated that the blue fox's outbreak of abortion events was caused by brucellosis via the B. melitensis strain. Then 726 serum samples were tested by RBPT and SAT to determine the prevalence of brucellosis on the farm. A comprehensive epidemiological and reproductive status survey of the infected blue fox population was performed. The seropositive rate was found to be 67.90% (493/726) by RBPT and 41.32% (300/726) by SAT. The technicians had stopped feeding the foxes with chicken carcasses and instead fed them raw ground sheep organs (lungs, tracheae, placentae, and dead sheep fetuses) infected by B. meliteneis strains, and that this change in diet caused the outbreak of abortion events. The high abortion rate (55%) and low cub survival rate (65%) were the most distinctive features of the outbreak; these factors led to severe economic losses. Feeding cooked sheep/goat offal and strict breeding management is necessary for disease prevention.

Chromosomal Instability at Fragile Sites in Blue Foxes, Silver Foxes, and Their Interspecific Hybrids

A cytogenetic assay based on fragile sites (FS) enables the identification of breaks, chromatid gaps, and deletions. In healthy individuals, the number of these instabilities remains low. Genome stability in these species is affected by Robertsonian translocations in the karyotype of the blue fox and by B chromosomes in the silver fox. The aims of the study were to characterise the karyotype of blue foxes, silver foxes, and their hybrids and to identify chromosomal fragile sites used to evaluate genome stability. The diploid number of A chromosomes in blue foxes ranged from 48 to 50, while the number of B chromosomes in silver foxes varied from one to four, with a constant number of A chromosomes (2n = 34). In interspecific hybrids, both types of karyotypic variation were identified, with the diploid number of A chromosomes ranging from 40 to 44 and the number of B chromosomes varying from 0 to 3. The mean frequency of FS in foxes was 4.06 ± 0.19: 4.61 ± 0.37 in blue foxes, 3.46 ± 0.28 in silver foxes, and 4.12 ± 0.22 in hybrids. A relationship was identified between an increased number of A chromosomes in the karyotype of the hybrids and the frequency of chromosomal breaks. The FS assay was used as a biomarker for the evaluation of genomic stability in the animals in the study.

Survival of SARS-CoV-2 on Clothing Materials

In order to plan and execute proper preventative measures against COVID-19, we need to understand how SARS-CoV-2 is transmitted. It has been shown to remain infectious on surfaces from hours to days depending on surface type and environmental factors. The possibility of transmission through fur animals and contaminated pelts, along with the safety of those working with them, is a major concern. SARS-CoV-2 can infect minks and raccoon dogs and has spread to mink farms in numerous countries. Here, we studied the stability of SARS-CoV-2 on blue fox, Finn raccoon, and American mink pelt, fake fur, cotton, plastic, faux leather, and polyester and tested its inactivation by UV light and heat treatment. We detected infectious virus up to 5 days on plastic, up to 1 day on fake fur, less than a day on cotton, polyester, and faux leather, and even 10 days on mink fur. UV light failed to inactivate SARS-CoV-2 on pelts, most likely due to the mechanical protection by the fur. Hence, it should not be used to inactivate the virus on fur products, and its use for other surfaces should also be considered carefully. Heat treatment at 60°C for 1 h inactivated the virus on all surfaces and is a promising method to be applied in practice. This study helps prevent further spread of COVID-19 by increasing our understanding about risks of SARS-CoV-2 spread through contaminated clothing materials and giving important information needed to improve safety of those working in the production line as well as people using the products.

Declined expressing mRNA of beta-defensin 108 from epididymis is associated with decreased sperm motility in blue fox (Vulpes lagopus)

Background Fecundity is important for farm blue fox (Vulpes lagopus), who with asthenospermia have be a problem in some of farms in China. A key symptom of asthenospermia is decreased sperm motility. The decreased secreting beta-defensin108 (vBD108) of blue fox is speculated be related to asthenospermia. To clarify this idea, the mRNA expression of vBD108 in testis and epididymis of blue foxes with asthenospermia were detected and compared to the healthy one. The antibody was prepared and analyzed by immunohistochemistry. Results The vBD108 in testis and epididymis was found both in blue fox with asthenospermia and healthy group by the method of immunohistochemistry. The expression of vBD108 mRNA in testes (P < 0.05) and epididymal corpus (P < 0.0001) in asthenospermia group was lower than that in healthy group. Conclusions These results suggested that vBD108 deficiency may related to blue fox asthenospermia. Meanwhile, the study on the blue fox vBD108 provides a hopeful direction to explore the pathogenesis of blue fox asthenospermia in the future.

Spleen arterial vascularization in fur-bearing animals in cages

The spleen is an unpaired parenchymal organ located in the abdominal cavity, performing immunological, filtration and hematopoietic functions and, in connection with the listed functional features, plays an essential role in the organism. Authors obtained detailed information on the arterial vascularization of the spleen of fur-bearing animals of Carnivora, since the information available in the special literature on the morphological aspects of the blood supply of the named organ is fragmentary. A comprehensive study of the spleen was carried out, which made it possible to confirm the general morphological patterns, to reveal the characteristic specific and intraspecific features of the vascularization of the spleen in fur-bearing animals in cages. The new data obtained on the morphology of the vessels vascularizing the spleen are original and give a complete picture of the organ. Analyzing the material obtained from own research, and comparing it with information from special literature, allows to assert that the blood supply to the spleen in fur-bearing animals of cellular content (silver-black fox, blue fox, sable and American mink) is carried out through the splenic artery, which also gives off branches to the stomach, pancreas and great omentum. If in the methods of branching of the splenic artery in the studied animals we note a great similarity, then in the number of branches branching from it there are significant differences. As a result of the study, it was found that the spleen is vascularized by the splenic artery extending from the celiac artery.

Morphometric Characteristics of the Spermatozoa of Blue Fox (Alopex lagopus) and Silver Fox (Vulpes vulpes)

The results presented in this study are the first such extensive characterization of the sperm morphometry of the blue fox (Alopex lagopus) and silver fox (Vulpes vulpes), as representatives of the family Canidae. Canine spermatozoa, especially the sperm of farmed foxes, are not often described in studies on reproduction. The aim of the study was a detailed comparison of the morphometric dimensions and shape of the sperm of two fox species: silver fox and blue fox. Semen collected from 10 silver foxes and 10 blue foxes was used for the study. The specimens were stained with silver nitrate. Measurements were performed of the length, width, perimeter, and area of the head; the area of the acrosome and its coverage; the length of the midpiece and its coverage; the length of the tail; and the length of the end piece of the tail. In addition, four head shape indices were calculated: ellipticity, elongation, roughness and regularity. The following values for the morphometric parameters and shape indices were obtained for blue fox and silver fox, respectively: head length—6.72 µm and 6.33 µm; head width—4.54.µm and 4.21 µm; head perimeter—18.11 µm and 17.37 µm; head area—21.94 µm2 and 21.11 µm2; acrosome area—11.50 µm2 and 10.92 µm2; midpiece length—12.85 µm and 12.79 µm; tail end piece length—3.44 µm and 3.28 µm; tail length—65.23 µm and 65.09 µm; acrosome coverage—52.43% and 52.83%; midpiece coverage—19.71% and 19.65%; sperm length—71.95 µm and 71.42 µm; ellipticity—1.49 and 1.52; elongation—0.19 and 0.20; roughness—0.84 and 1.88; regularity—1.09 and 0.99. The significance of differences between species was verified by Tukey’s test at p ≤ 0.05. Statistically significant differences between species were found for the following parameters: head length, width, perimeter and area; acrosome area; tail, end piece, and total sperm length; roughness and regularity. The differences in the size and shape of sperm can be used to establish reference patterns for fox sperm enabling more accurate species identification.